JP2011505404A - ヒトcd154−結合性の合成ペプチドおよびその使用 - Google Patents
ヒトcd154−結合性の合成ペプチドおよびその使用 Download PDFInfo
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Abstract
Description
[式中、LはLeuであり、PはProであり、TはThrであり、RはArgであり、HはHisであり、MはMetであり、AはAlaであり、GはGlyであり、およびKはLysである]。
この研究を通して、7個のアミノ酸残基とジスルフィド結合による環化を可能にする両端で側面に位置する2個のシステインを含むペプチド配列パネルから成り、M13ファージで発現され、キャプシドpIIIファージタンパク質と遺伝子工学的に融合され、そのファージ表面に無作為に発現される、ペプチドライブラリー(ペプチドの多様性<109)を用いた。このファージライブラリーを、既知のモデル(Hetian L. et al, J. Biol. Chem. 2002)から本発明者らにより発展されたバイオパニング・インビトロ・モデルによってスクリーニングした。ファージ(200μlのTBS中、1×1011CFU)を、ヒト組み換えCD154と一緒に室温で1時間インキュベートして、その後プレートに撒いた。このファージをTBS−Tを用いて数回洗浄してCD154と結合していない非特異的なファージを取り除き、CD154と結合したファージを、100μlの0.2M グリシン、pH2.2を用いて溶出することにより回収し、15μlの1M Tris−HCl、pH9.1を用いて10分間中和した。ファージ数は、溶出液の段階希釈物をLB寒天培地(アガロース7g/リッター、MgCl2・6H2O 1g;Sigma)上の宿主のテトラサイクリン耐性の大腸菌ER2738細胞(New England Biolabs, Hitchin, U.K.)に加え、テトラサイクリン(Kramel Biotech, Cramlington, U.K.)存在下のIPTG/X−Gal LB寒天培地に撒いて、滴定することにより評価した。
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Claims (29)
- CD154受容体の活性部位に選択的に結合することができ、CD40:CD154相互作用を阻害することができるペプチドであって、配列番号:13で示されるCD154結合性のエプタ−アミノ酸配列を含む、ペプチド。
- 7アミノ酸長から30アミノ酸長である、請求項1記載のペプチド。
- 配列番号:13で示されるCD154結合性のアミノ酸配列が、該配列の両端の側面に2個のシステインをさらに含み、それにより配列番号:6で示されるノナ−アミノ酸配列を提供することとなる、請求項1または2記載のペプチド。
- 配列番号:6で示されるアミノ酸配列からなる、請求項3記載のペプチド。
- 環状である、請求項3または4記載のペプチド。
- 請求項1〜5のいずれか一項記載のペプチドの複数のコピーを含む、多量体構造物。
- 請求項1〜5のいずれか一項記載のペプチドの各コピーが、直鎖または環状のいずれかである、請求項6記載の多量体構造物。
- 請求項1〜5のいずれか一項記載のペプチドの各コピーが、少なくとも1つの他のコピーと少なくとも1つのアミノ酸スペーサーにより連結されている、請求項6または7記載の多量体構造物。
- アミノ酸スペーサーが、Gly(G)残基である、請求項8記載の多量体構造物。
- ペプチドの各コピーが、直接または間接的に連結されるアミノ酸コアを含む、請求項8または9記載の多量体構造物。
- アミノ酸コアが、複数のLys(K)残基からなる、請求項10記載の多量体構造物。
- ペプチドが、配列番号:13で示される直鎖のエプタ−アミノ酸配列からなる、請求項6〜11のいずれか一項記載の多量体構造物。
- 請求項1〜5のいずれか一項記載の少なくとも1つのペプチド、または請求項6〜13のいずれか一項記載の少なくとも1つの多量体構造物を含み、生体分子、診断剤および治療剤からなる群より選択される分子と複合体化された、コンジュゲート。
- 治療剤が、抗炎症性剤、免疫抑制剤、免疫調節剤または抗腫瘍剤である、請求項14記載のコンジュゲート。
- 治療剤が、活性化内皮細胞、腫瘍細胞または活性化によりその表面にCD154を発現する細胞に対して細胞傷害効果を発揮することができる、請求項14記載のコンジュゲート。
- 診断剤が、インビボで検出可能な分子である、請求項14記載のコンジュゲート。
- 診断剤が、インビトロアッセイのための検出可能な分子である、請求項14記載のコンジュゲート。
- 医薬としての、請求項1〜5のいずれか一項記載のペプチド、または請求項6〜13のいずれか一項記載の多量体構造物。
- CD40:CD154相互作用に関係する疾患または障害の処置のための医薬を製造するための、請求項1〜5のいずれか一項記載のペプチドまたは請求項6〜13のいずれか一項記載の多量体構造物の使用。
- 疾患または障害が、炎症である、請求項20記載の使用。
- 炎症が、アテローム性動脈硬化症、自己免疫疾患または移植による拒絶反応から選択される炎症性疾患である、請求項21記載の使用。
- 自己免疫疾患が、全身性エリテマトーデス(SLE)、リウマチ性関節炎(RA)、クローン病または乾癬である、請求項22記載の使用。
- 炎症が、関節炎を伴う炎症、接触皮膚炎、高IgE症候群、炎症性腸疾患、アレルギー性喘息を含むアレルギーおよび特発性炎症性疾患からなる群より選択される、請求項21記載の使用。
- 移植による拒絶反応が、移植された膵臓の膵島細胞、皮膚、骨髄、肝臓、心臓および腎臓を含む、請求項22記載の使用。
- 疾患が、腫瘍性疾患である、請求項20記載の使用。
- 腫瘍性疾患が、慢性リンパ性白血病(CLL)である、請求項26記載の使用。
- 疾患が、移植片対宿主病である、請求項20記載の使用。
- 請求項1〜5のいずれか一項記載のペプチドまたは請求項6〜13のいずれか一項記載の多量体構造物の有効量、および医薬上許容されるビヒクルおよび/または賦形剤を含む、医薬組成物。
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EP07122164A EP2067785A1 (en) | 2007-12-03 | 2007-12-03 | Human CD154-binding synthetic peptide and uses thereof |
PCT/EP2008/066349 WO2009071486A1 (en) | 2007-12-03 | 2008-11-27 | Human cd154-binding synthetic peptide and uses thereof |
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EP2310405A4 (en) | 2008-07-11 | 2011-08-10 | Phylogica Ltd | PEPTIDE INHIBITORS OF SIGNALING USING CD40L AND USES THEREOF |
EP2569418B1 (en) | 2010-05-12 | 2016-11-02 | RegenMed (Cayman) Ltd. | Bioactive renal cells |
AU2019217207A1 (en) | 2018-02-12 | 2020-08-27 | Diabetes-Free, Inc. | Improved antagonistic anti-human CD40 monoclonal antibodies |
WO2020102454A1 (en) | 2018-11-13 | 2020-05-22 | Regents Of The University Of Minnesota | Cd40 targeted peptides and uses thereof |
CN115845063A (zh) * | 2022-12-20 | 2023-03-28 | 中国医学科学院医学生物学研究所 | 与急性t淋巴细胞白血病治疗相关的靶点cd40lg及其应用 |
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WO2002018446A2 (en) * | 2000-09-01 | 2002-03-07 | Biogen, Inc. | Methods of designing and producing compounds having improved binding affinity for cd154 or other trimeric proteins |
JP2004503507A (ja) * | 2000-06-09 | 2004-02-05 | ブリストル−マイヤーズ スクイブ カンパニー | リンパ球シグナルのブロックおよびlfa−1媒介性接着のブロックによる細胞性免疫応答の調節方法 |
JP2004517814A (ja) * | 2000-09-01 | 2004-06-17 | バイオジェン インコーポレイテッド | モノクローナル抗体5c8およびCD154の共晶構造、ならびに薬物設計におけるその使用 |
WO2007033058A2 (en) * | 2005-09-13 | 2007-03-22 | Trustees Of Dartmouth College | Compositions and methods for regulating rna translation via cd154 ca-dinucleotide repeat |
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WO2005003175A2 (en) | 2003-06-13 | 2005-01-13 | Biogen Idec Ma Inc. | Aglycosyl anti-cd154 (cd40 ligand) antibodies and uses thereof |
US8647625B2 (en) | 2004-07-26 | 2014-02-11 | Biogen Idec Ma Inc. | Anti-CD154 antibodies |
CN100369932C (zh) * | 2005-04-07 | 2008-02-20 | 苏州大学 | 抗人cd154单克隆抗体及其应用 |
WO2009137471A2 (en) * | 2008-05-05 | 2009-11-12 | University Of Miami | Azo dye related small molecule modulators of protein-protein interactions |
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JP2004503507A (ja) * | 2000-06-09 | 2004-02-05 | ブリストル−マイヤーズ スクイブ カンパニー | リンパ球シグナルのブロックおよびlfa−1媒介性接着のブロックによる細胞性免疫応答の調節方法 |
WO2002018446A2 (en) * | 2000-09-01 | 2002-03-07 | Biogen, Inc. | Methods of designing and producing compounds having improved binding affinity for cd154 or other trimeric proteins |
JP2004517814A (ja) * | 2000-09-01 | 2004-06-17 | バイオジェン インコーポレイテッド | モノクローナル抗体5c8およびCD154の共晶構造、ならびに薬物設計におけるその使用 |
WO2007033058A2 (en) * | 2005-09-13 | 2007-03-22 | Trustees Of Dartmouth College | Compositions and methods for regulating rna translation via cd154 ca-dinucleotide repeat |
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JPN5010015343; MODERN RHEUMATOLOGY Vol.15, No.6, 20051201, P.423-426 * |
JPN5010015344; ANNUAL REVIEW OF IMMUNOLOGY Vol.14, 199601, P.591-617 * |
JPN5010015345; JOURNAL OF INVESTIGATIVE DERMATOLOGY Vol.126, No.1, 200601, P.105-113 * |
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US20110144038A1 (en) | 2011-06-16 |
CN101883780A (zh) | 2010-11-10 |
WO2009071486A1 (en) | 2009-06-11 |
ES2436427T3 (es) | 2014-01-02 |
JP5593231B2 (ja) | 2014-09-17 |
CN101883780B (zh) | 2015-04-22 |
EP2222692B1 (en) | 2013-08-28 |
PL2222692T3 (pl) | 2014-01-31 |
US8507448B2 (en) | 2013-08-13 |
EP2067785A1 (en) | 2009-06-10 |
EP2222692A1 (en) | 2010-09-01 |
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